间充质干细胞在银屑病中的应用

【摘要】 间充质干细胞是干细胞家族的重要成员,具有自我更新、多向分化潜能和免疫调节作用及低免疫原性等特点。有文献报告间充质干细胞用于治疗银屑病获得不同程度的疗效,且有较长的疾病缓解期。本文综述间充质干细胞在银屑病中的应用及相关研究进展。     

骨髓间充质干细胞在皮肤疾病治疗中的研究进展

骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BM-MSC)是一类来源于骨髓的非造血干细胞,它具有多向分化潜能和免疫调节作用,因为其获取容易,并能在体外扩增,使其成为应用最广泛的间充质干细胞。目前骨髓间充质干细胞在皮肤病的研究中受到广泛的关注,包括伤口愈合、放射性皮炎、系统性红斑狼疮(SLE)、硬皮病和银屑病等。     

银屑病患者骨髓间充质干细胞体外自发向血管内皮细胞分化1例

目的:揭示1例银屑病患者骨髓间充质干细胞体外自发向血管内皮细胞分化.方法:采用密度梯度离心法分离骨髓单一核细胞,通过贴壁法培养骨髓间充质干细胞,经全量换液和2次半量换液后(即培养9 d)流式细胞术鉴定正常对照细胞免疫表型,再培养2 d,采用流式细胞术和Dil-ac-LDL、FITC-UEA-1双荧光染色法,分别对患者的细胞和正常对照细胞进行内皮细胞鉴定.结果:患者细胞在培养11 d后大部分分化为血管内皮细胞,而正常对照则仍为较纯的间充质干细胞.结论:该患者骨髓间充质干细胞体外培养自发向血管内皮细胞分化.这一现象表明银屑病患者骨髓间充质干细胞向血管内皮细胞分化过程中可能存在异常,其间充质干细胞有易于向血管内皮细胞分化的倾向.     

胎牛血清对骨髓间充质干细胞增殖的影响

目的揭示不同来源胎牛血清对骨髓间充质干细胞增殖的影响。方法采用密度梯度离心法分离骨髓单一核细胞,通过贴壁法分别使用2种不同来源的胎牛血清培养银屑病患者和正常对照者骨髓间充质干细胞,记录各组细胞传1代所需的时间;MTT比色法检测患者骨髓间充质干细胞的增殖活性;An-nexinV法检测银屑病患者骨髓间充质干细胞自发凋亡率。结果使用胎牛血清1,银屑病患者骨髓间充质干细胞在培养过程中逐渐出现凋亡,最后全部死亡,正常对照者骨髓间充质干细胞传代所需时间也较长;而使用胎牛血清2,患者细胞可以传代,正常对照细胞传代所需时间缩短(P0.05)。结论胎牛血清是影响骨髓间充质干细胞增殖的重要因素。     

银屑病患者皮损间充质干细胞IGFBP3和TNFSF15 mRNA的表达

目的探讨胰岛素样生长因子结合蛋白-3(IGFBP3)和肿瘤坏死因子超家族成员-15(TNFSF15)在银屑病发生发展中的作用及临床意义。方法分离扩增15例银屑病患者皮损及15例正常人皮肤的间充质干细胞,采用实时荧光定量聚合酶链反应(RT-PCR)分别检测IGFBP3和TNFSF15的m RNA表达水平。结果银屑病患者皮肤间充质干细胞中IGFBP3和TNFSF15 m RNA表达水平较对照组明显降低,且差异具有统计学意义(P0.05)。结论银屑病患者皮肤间充质干细胞IGFBP3 m RNA表达降低可能与银屑病角质形成细胞增殖活性增高及皮损局部血管形成增加有关。而TNFSF15 m RNA表达降低可能与银屑病皮损处血管内皮细胞的增殖有关。     

银屑病患者骨髓间充质干细胞表面标志表达情况研究

目的研究银屑病患者骨髓间充质干细胞的体外分离培养及表面标志的表达情况,为银屑病患者间充质干细胞特性研究提供了基础。方法采用密度梯度离心法分离患者与对照组骨髓单一核细胞,通过贴壁法培养骨髓间充质干细胞,用流式细胞术鉴定细胞表面标志CD44、CD29、CD34、CD45、HLA-DR。结果经流式细胞仪测定的第三代培养的BMSCs表面标志CD44、CD29阳性表达,CD34、CD45、HLA-DR阴性表达。分析CD44、CD29,发现银屑病组明显低于对照组。结论密度梯度离心法结合贴壁法可获得较纯的BMSCs。银屑病患者表面标志CD44、CD29与正常人比较低表达。     

间充质干细胞移植治疗自身免疫性疾病的进展

间充质干细胞(MSC)是来源于发育早期中胚层的成体干细胞,具有高度增殖、自我更新能力、多向分化潜能、损伤组织修复功能和免疫抑制能力.该文综述间充质干细胞,主要是脐带间充质干细胞(hUMSCs)和骨髓间充质干细胞(BMMSC),用于移植治疗系统性红斑狼疮(SLE)、类风湿关节炎(RA)、硬皮病(SSc)、皮肌炎(DM)的研究进展,并显示间充质干细胞移植(MSCT)在未来的分子治疗中是非常有前景的一种方法.     

银屑病骨髓间充质干细胞分泌白介素-6和血管内皮生长因子的检测

目的:研究银屑病患者骨髓间充质干细胞(MSCs)细胞因子分泌水平及其与病情活动的相关性,进一步探讨银屑病的发病机制.方法:采用密度梯度离心分离和贴壁培养法对24例银屑病患者和20例正常对照骨髓MSCs进行分离培养,采用ELISA方法测定MSCs培养上清白介素(IL)-6和血管内皮生长因子(VEGF)的表达.结果:银屑病患者骨髓MSCs培养上清液IL-6浓度较正常人明显增高(P＜0.01),VEGF浓度较正常人明显降低(P＜0.01).患者IL-6及VEGF水平与银屑病皮损面积及严重度指数(PASI)评分无相关性(P＞0.05).结论:银屑病患者骨髓MSCs细胞因子分泌异常,银屑病患者骨髓造血微环境有异常,进而对银屑病造血干细胞发育可能产生影响.     

间充质干细胞移植治疗系统性红斑狼疮的研究进展

间充质干细胞是来源于发育早期中胚层的成体干细胞,具有高度增殖、自我更新能力、多向分化潜能、损伤组织修复功能和免疫抑制能力。该文综述间充质干细胞,主要是脐带间充质干细胞和骨髓间充质干细胞用于治疗系统性红斑狼疮的研究进展,并提示间充质干细胞移植在未来的细胞治疗中是非常有前景的一种方法。     

骨髓间充质干细胞治疗吸入性肺损伤研究进展

成体干细胞来源广泛,无伦理争议,成为近几年关注热点.研究表明以骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)为代表的成体干细胞具有较强的多系分化潜能.骨髓间充质干细胞在肺损伤状态下可以向肺部移行,表现出肺上皮细胞的表型,参与损伤肺组织的修复重建,并可通过旁分泌途径等减轻肺损伤.并有研究表明干细胞的植入可以缓解和抑制损伤肺的病理进程.因此BMSCs移植为吸入性肺损伤的防治提供了一种崭新有效的方法.本文就BMSCs治疗热烟雾吸入性肺损伤中的应用综述如下.     

Biological characteristics and gene expression pattern of bone marrow mesenchymal stem cells in patients with psoriasis

Mesenchymal stem cells (MSCs) have immunoregulatory and proangiogenic effects and are suggested to be involved in the pathological processes of immune‐related diseases, including psoriasis. Biological characteristics of bone marrow MSCs (BMSCs) from patients with autoimmune diseases, such as systemic lupus erythematosus or rheumatoid arthritis, but not psoriasis, have been characterized. We compared the gene expression profile and biological characteristics of BMSCs from patients with psoriasis and healthy controls. Although the phenotype, differentiation potential and ability to support CD34⁺ cell proliferation were similar to those of normal BMSCs, psoriatic BMSCs showed aberrant proliferative activity, increased apoptosis rate and a characteristic gene expression profile. These aberrations may develop after the abnormal immune response in psoriasis and result in BMSC dysfunction. The functionally deficient BMSCs may then fail to suppress overactive immune cells, thereby contributing to the pathogenesis of psoriasis.     

Mesenchymal stem cells for the treatment of psoriasis: a comprehensive review

Mesenchymal stem cells (MSCs) have recently been shown to have not only regenerative capabilities but also immunomodulating properties. For this reason, they are currently under investigation in clinical trials for the treatment of several autoimmune systemic disorders. Psoriasis is a systemic immune-mediated disease for which MSCs could have therapeutic potential. We analysed the existing literature with regard to MSC-based strategies for the treatment of psoriasis, using the MEDLINE, Embase, Scopus and Cochrane Library electronic databases from inception to the date of study. A number of studies confirm the involvement of MSCs in psoriasis pathogenesis and therefore designate MSCs as an important potential therapeutic tool in this setting. Preclinical data are mostly based on imiquimod-induced murine models of psoriasis, and confirm the anti-inflammatory and immunomodulatory action of MSCs in the setting of psoriasis. Six patients affected by psoriasis were described in four clinical studies. Despite significant differences in terms of therapeutic protocols and clinical outcomes, the MSC-based regimens were efficacious in 100% of the cases. Despite more data still being needed, MSCs could be a promising therapy for psoriasis.     

The mesenchymal stem cell profile in psoriasis

Background The expression of inducible nitric oxide synthase (iNOS) and vascular endothelial growth factor (VEGF) and the level of total oxyradical scavenging capacity have been evaluated extensively in the cutaneous cells of patients with psoriasis. As yet, no indications are available about the undifferentiated cells, the mesenchymal stem cells (MSCs), isolated from skin. Objectives To isolate MSCs in patients with psoriasis and to compare them with those obtained from atopic and healthy subjects, in order to analyse whether MSCs show some typical psoriatic profiles and to understand whether pathophysiological events leading to psoriasis start early at the stem cell level. Methods MSCs isolated from seven patients with psoriasis, seven patients with acute atopic dermatitis and seven healthy subjects were characterized by fluorescence‐activated cell sorting analysis. VEGF and nitric oxide (NO) content was measured in conditioned medium, the expression of VEGF and iNOS was analysed by immunohistochemistry, and the total oxyradical scavenging capacity towards peroxynitrite was tested. Results VEGF content was highest in the medium conditioned by psoriatic perilesional MSCs, whereas NO concentration was maximally increased in medium conditioned by MSCs isolated from lesional psoriatic skin. The ability to neutralize the oxidizing effects of peroxynitrite was lower for MSCs isolated from lesional psoriatic skin compared with other MSCs, except for MSCs of lesional atopic skin. Conclusions The microenvironment in psoriasis differs from those of atopic dermatitis and healthy skin; it could induce resident MSCs to produce angiogenic and proinflammatory mediators which lead to a reduction in the antioxidant capacity of these cells, contributing to the development of skin lesions in psoriasis.     

Mesenchymal stem cells in psoriatic lesions affect the skin microenvironment through circular RNA

Psoriasis is an autoimmune skin disease. Our previous studies revealed abnormal immune regulation of skin mesenchymal stem cells (S‐MSCs) in psoriatic lesions. Circular RNA (circRNA) molecules were recently discovered as a new class of non‐coding regulatory RNAs. Their role in the pathogenesis of psoriasis has not yet been studied. To explore potential circRNA‐mediated mechanisms of S‐MSCs in the pathogenesis of psoriasis, we sequenced mRNAs and circRNAs of MSCs from normal skin and psoriatic lesions, followed by functional prediction and interaction analyses. In total, 129 circRNAs were differentially expressed, including 123 up‐regulated and 6 down‐regulated circRNAs, in MSCs from psoriatic lesions. Pathway analysis showed that the genes significantly down‐regulated in psoriatic as compared to normal S‐MSCs were mainly involved in JAK‐STAT signalling. According to a circRNA‐miRNA‐mRNA interaction network, the expression of circRNAs associated with these mRNAs was also down‐regulated in MSCs of psoriatic skin lesions. Knockdown of the circRNA gene chr2:206992521|206994966 reduced the capacity of S‐MSCs to inhibit T‐cell proliferation upon co‐culture in normal as well as lesion‐derived S‐MSCs. Secreted‐cytokine profiles (IL‐6, IL‐11 and hepatocyte growth factor) were also similar in normal and lesion‐derived S‐MSCs after circRNA knockdown. Thus, the circRNA chr2:206992521|206994966 in S‐MSCs from psoriatic lesions affects the activity of T lymphocytes in local lesions by influencing their cytokine secretion. Taken together, our findings indicate that circRNA mediates the role of S‐MSCs in the pathogenesis of psoriasis.     

间充质干细胞治疗结核分枝杆菌感染的基础研究进展

 间充质干细胞（MSCs）是一群具有高度自我更新能力和多向分化潜能的多能干细胞，已有多项研究证明其对结核分枝杆菌感染的治疗潜力。MSCs可通过抗菌肽与关键酶的分泌、细胞表面受体的表达，直接或间接地介导受宿主细胞由免疫抑制状态向激活状态转化，进而作为细胞治疗的一种手段，在足量抗菌药物覆盖的基础上，辅助杀灭以结核分枝杆菌为主的分枝杆菌感染，并抑制过度的炎症反应，减少不必要的组织损伤。本文就间充质干细胞治疗结核分枝杆菌感染的基础及临床前研究进展作一概述。     

Effect of biologic therapies targeting tumour necrosis factor-α on cutaneous mesenchymal stem cells in psoriasis

Summary Background Psoriasis is a Th1 immune‐mediated, inflammatory disease, in which skin lesions appear many years before the related metabolic and cardiovascular comorbidities, according to the theory of the ‘psoriatic march’. Inducible nitric oxide synthetase (iNOS), tumour necrosis factor (TNF)‐α and vascular endothelial growth factor (VEGF) are directly implicated in determining both skin lesions and systemic involvement in psoriasis. Reactive oxygen species actively promote the secretion of inflammatory Th1 cytokines directly involved in the pathogenesis of psoriasis. Objectives Evaluation of VEGF expression and production, nitric oxide (NO) production, iNOS expression, and the antioxidant response of mesenchymal stem cells (MSCs), both before and after 12 weeks of treatment with the TNF‐α inhibitors adalimumab or etanercept. Methods Biochemical, morphological and immunohistochemical analyses were performed in MSCs isolated from nonlesional, perilesional and lesional skin of patients with psoriasis, before and after treatment. Results The treatments were able to reduce the expression and production of VEGF, the expression of iNOS and the production of NO in MSCs of patients with psoriasis. TNF‐α inhibitors also reduced the oxidative damage in MSC membrane and proteins, several antioxidant systems responded to treatments with a general inhibition of activities (glutathione S‐transferase and catalase) and these effects were also supported by a general decrease of total oxyradical scavenging capacity towards hydroxyl radicals and peroxynitrite. Conclusions TNF‐α inhibitors are able to change the physiopathological pathway of psoriasis, and our results suggest their therapeutic effects already take place at the level of MSCs, which probably represent the cells primarily involved in the ‘psoriatic march’.     

Cutaneous mesenchymal stem cells: status of current knowledge,implications for dermatopathology

Stem cell biology is currently making its impact on medicine, which will probably increase over the next decades. It not only influences our therapeutic thinking caused by the enormous plasticity of stem cells but also affects diagnostic and conceptual aspects of dermatopathology. Although our knowledge of the keratinocytic stem cells located within the follicular bulge has exploded exponentially since their discovery in 1990, the concept of cutaneous mesenchymal stem cells (MSCs) is new. Described initially in 2001 in mice, MSCs later were also found in the human dermis. The connective tissue sheath and the papilla of the hair follicle probably represent the anatomical niche for cutaneous MSCs. In line with the cancer stem cell hypothesis, mutations of these cells may be the underlying basis of mesenchymal skin neoplasms, such as dermatofibrosarcoma protuberans. Furthermore, research on cutaneous MSCs may impact our thinking on the interaction of the epithelial component of skin neoplasms with their surrounding stroma. We are only in the early stages to recognize the importance of the potential contributions of cutaneous MSC research to dermatopathology, but it is not inconceivable to assume that they could be tremendous, paralleling the early discovery of the follicular bulge as a stem cell niche. Sellheyer K, Krahl D. Cutaneous mesenchymal stem cells: status of current knowledge, implications for dermatopathology.     

Mesenchymal stem cell therapy in skin: why and what for?

In recent years, few stem cells have gained as much clinical notoriety as mesenchymal stem cells. Indeed, MSCs are already in use for a range of systemic inflammatory and autoimmune conditions that also affect the skin, such as acute and chronic graft versus host disease or lupus erythematosus. Most interestingly, these cells are able to improve skin wound healing in multiple preclinical models and few patient series. An additional potential of these cells is the delivery of missing structural elements in skin inherited disorders. However, we here contend that MSCs are not appropriate for cell replacement therapies in the context of wound healing. Indeed, engraftment of cells in the dermis is poor in the absence of irradiation and the observed effects seem mainly due to paracrine factors. In this viewpoint, we favour the hypothesis of a replete niche and competition with resident mesenchymal populations in the dermis not allowing the engraftment of newly delivered MSCs. Consequently, we propose that the benefit of MSCs may be at least in part reproduced by the growth factors or immunomodulatory molecules that they produce. In any case, the rapid progress in this field has allowed the emergence of important questions in skin biology that need to be addressed in parallel with the predictable future use of MSCs in the clinic.     

Mesenchymal stem cell therapies: the quest for fine‐tuning

Mesenchymal stem cells (MSCs) have a vastly unharnessed therapeutic potential with close to 400 studies currently registered on clinicaltrials.gov for evaluation of their clinical promises. While many of these investigations are for immune‐mediated disorders, there is no established consensus on how to optimize the immunomodulatory properties of MSCs. Factors that could be used to predict efficacy of MSC therapies include donor heterogeneity, recipient environment and drug interactions. Incorporating pertinent quality control parameters to maximize the clinical potential of MSCs through good manufacturing practice (GMP) production of clinical grade cells could lead to the realization of greater therapeutic success.